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Работа фрилансера в журнале

Многие фрилансеры имеют опыт работы в различных изданиях, журналистское, филологическое образование. Найти журналиста в каталоге FreelanceJob. Юлия Куликова. Екатерина Храмова. Мария Курляндцева. Александр Чивилев. Екатерина Висицкая. Дарья Макарова. Валерия Маршевская. Алина Мурашкина. Ekaterina Milenkovic Baleva. Татьяна Лесникова. Наталья Ивановская. Александр Власов. Евгения Григорова Морозова. Валерия Денисова. Дмитрий Ефремов.

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Вы сами назначаете стоимость за свою работу. А именно, текст пишется полностью своими словами без смены основного смысла. У данной услуги есть свои приемы, а именно:. Ведь основным критерием в выборе исполнителя заказа, является его высокий рейтинг, хороший показатель КПД и конечно же высокая обязательность. Лишь добившись максимальных показателей, можно рассчитывать на персональные заказы и достойную оплату своего труда в интернете. Для журналиста работа рерайтера — это гораздо более простая, чем создание новых уникальных текстов.

Она отлично подходит для старта карьеры в сети Интернет. Главные требования заказчика — грамотность и профессионализм при написании материалов. Информации нужно много, как правило более текстов. Нередко это циклы статей с одной тематикой. Стоимость подобных заказов оговаривается индивидуально. Получить эту работу довольно сложно, ведь на нее, как правило, очень много желающих фрилансеров, а конкуренция просто сумасшедшая.

Но постоянно просматривая многочисленные объявления, есть вероятность отыскать этот приятный и весьма доходный от до у. Внештатный сотрудник в интернет-журнал. Мечта любого журналиста, а не только фрилансера. Ведь получить работу в подобном журнале сравнимо с выигрышем в лотерею. На каждом подобном сайте уже имеется полностью сформированный штат квалифицированных журналистов.

И поверьте, они дорожат своим местом работы. Интернет—журналы, имеющие свободную регистрацию, можно встретить весьма нечасто. Женские интернет-журналы — наиболее реальный работодатель для журналиста. Если же вам удалось устроиться на данную работу, считайте, что счастливый билет достался именно вам.

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Образование журналиста сделает данную работу наиболее творческой. Колонка в определенное время публикуется в каком-либо издании. На сайте имеет свое постоянное место. Может функционировать, как отдельная независимая рубрика.

Подразделяются колонки на несколько видов:. А определение журналист-блоггер является наиболее весомым и достойным для данной работы в интернете. Необходимо лишь:.

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For quantification of uterine stromal areas and stromal vessel luminal area, hematoxylin—eosin HE and CD31 stained photomicrographs of KO mice were analyzed using ImageJ Image analysis software Rasband, W. Total RNA was extracted from mice uterine tissues by homogenizing the tissues in 1 mL TRIzol Life Technologies, San Francisco, CA followed by the addition of uL chloroform to the lysate for phase separation by centrifugation at 13, rpm for 15 minutes.

The RNA pellet was allowed to dry and then re-dissolved in nuclease free water. Following first-strand synthesis with random primers, second-strand synthesis was performed with dUTP for generating strand-specific sequencing libraries. Libraries were sequenced on an Illumina HiSeq with parameters set for high output, single-end chemistry, and bp sequencing.

Samples were multiplexed to six samples per lane, yielding an average of 37 million reads per sample. For each read, we trimmed the first 6 nucleotides and the last nucleotides at the point at which the Phred score of an examined base fell below 20 using in-house scripts.

If, after trimming, the read was shorter than 45 bp, the whole read was discarded. Trimmed reads were mapped to the mouse reference genome mm10 with a known transcriptome index UCSC Known Gene annotation with Tophat v2. In these results, only the reads that mapped to a single genomic unique location, with a maximum of two mismatches in the anchor region of the spliced alignment, were reported.

The default settings for all other Tophat options were used. The program Cufflinks v2. Cuffdiff was used to obtain differential gene expression between the experimental groups using first-strand library type, providing gene model annotation and the genome sequence file for detection and correction of sequence-specific bias that can be caused by random hexamer during the process of library preparation.

We applied scRNA-seq using droplet microfluidics 10x Chromium on a single-cell suspension dissociated from E9. Nano-sized droplets that each contains a single cell with the bar-coded gel bead GEMs were generated using the Chromium controller 10x Genomics. Reverse transcription was performed with polyT primers containing cell-specific bar codes, unique molecular identifiers UMIs , and adaptor sequences.

All 10x libraries were sequenced in an Illumina HISeq instrument. We used the 10x Genomics Cell Ranger software v2. For graph-based clustering and differential gene expression analysis, we used Seurat 3. In Seurat, an initial filter was applied to select only the cells that had a minimum of unique transcripts; and to select only those genes that were expressed in at least 3 cells.

During normalization with sctransform, we also included in the model the mitochondrial mapping percentage as an unwanted source of variation. The expression of established lineage marker genes was used to assign cell types. Normally distributed data were analyzed using the Student unpaired two-tailed t test for the comparison of two groups, and one-way ANOVA with Tukey multiple comparison test for multiple group comparison.

If data were not normally distributed, or if distribution could not be determined due to small sample size, data were analyzed using a Mann-Whitney U test. For time to delivery Meier-Kaplan curves, differences between curves were evaluated by applying the log-rank test Kaplan-Meier log-rank value, p -value. A A schematic of the 5-FU—based submyeloablation protocol used for non-gonadotoxic BM transplantation. B Imaging using a fluorescent camera following dissection to separate the uterus U , placenta P , and embryo E on E9.

Right panel and middle panel are GFP fluorescence and X-ray images, respectively. They were analyzed by multicolor flow cytometry B. D-F Fluorescent images of trilineage differentiation of P-2 cultured BM cells grown in adipogenic media D , osteogenic media E or chondrogenic media F.

Nuclei are stained with DAPI blue. The bottom row for each panel is a higher magnification of the area in the middle row enclosed by a rectangle. Primary P-2 mouse uterine stromal cells served as positive control for decidualization. Values shown are expression levels relative to day 3. Results shown are the average of three independent experiments carried out in duplicates.

A Multicolor flow cytometry analysis of peripheral blood cells extracted from mice transplanted with BM from GFP donors following 5-FU submyeloablation. In the middle is the low-magnification image showing the mesometrial and antimesometrial sides of the implantation site. The mesometrial side is the side where the placenta, decidua basalis DB , and the major blood vessels are located. The mesometrial lymphoid aggregate MLAp is a transient structure between the myometrial layers that surrounds the radial branches of the uterine artery.

The antimesometrial side contains the rest of the maternal decidua in contact with the invading trophoblast. Red dashed line demarcates the giant cell GC layer. Maternal vascular spaces black dash have brown GFP-stained platelets black arrows and are interspersed between trophoblast cells and fetal vascular spaces green dash. Red arrows point to nucleated red blood cells characteristic of fetal vascular spaces. Red dashed line demarcates the GC layer.

Red arrows point to some decidual cells. A star demarcates the new lumen. GFP, green fluorescent protein. B Clustering of immune cells and DSCs following UMAP-based visualization of expression differences for cells using established lineage markers and C the same plot showing eGFP expression distribution within the same clusters. Percentages shown are of cells positive for the respective marker in each group. Bottom panel shows flow cytometry of dissociated uterine implantation site cells E9.

Numbers in each quadrant indicate mean percentage of cells. Multicolor flow cytometry was performed on peripheral blood and BM cells of nonpregnant and pregnant mice on E5. Cells from BM tibia or femur , spleen, peripheral blood, or implantation site of E9. A Live single cells of nonpregnant or pregnant E9.

Representative graphs are shown. Top panel shows multicolor flow cytometry gating strategy. Live leukocyte single cells were gated on CD The endometrial area in each photomicrograph is demarcated by a black dashed line. We would like to thank Prof. Gil Mor for his helpful insights and suggestions, Kristin Milano for technical assistance with immunostaining, and Gouzel Tokmoulina for technical assistance with flow cytometry. Abstract Decidua is a transient uterine tissue shared by mammals with hemochorial placenta and is essential for pregnancy.

Eaves, B. Introduction The decidua is a transient tissue lining the uterus of mammals with hemochorial placenta mice, humans, and numerous other mammalian species and is essential for pregnancy in these species. Download: PPT. Fig 1. Spatial and temporal contribution of BMDCs to decidua throughout mouse pregnancy.

Fig 2. Characterization of uterine BMDCs throughout pregnancy. Fig 4. Hoxaexpressing BMDCs induce expression of known decidualization genes in the implantation site to rescue pregnancy The maternal component of the implantation site consists of stromal cells of the uterine endometrium that undergo a tightly controlled process of decidualization upon contact with the early embryo.

Fig 5. Fig 6. BM transplantation from WT donors leads to stromal expansion and induces glandular formation in HoxAnull mice. Fig 7. BM transplantation from WT donors leads to decidualization reaction in Hoxanull mice. Discussion It is well established that uterine implantation sites are areas of infiltration of many BM-derived immune cells, which play important roles at the maternal—fetal interface to promote successful pregnancy reviewed in [ 6 ].

Animal studies Hoxa11 KO mice B6. F-actin staining F-actin staining of BM MSC cells and uterine cells was performed after 9 days in culture with different decidualization treatments. Image quantification and analysis For quantitation of proliferating GFP-positive and GFP-negative cells in the endometrial stromal or decidual compartments, 12 high-power confocal microscopy fields 4 high-power fields [HPFs] from each of 3 uterine sections per animal were assessed for each gestational time point.

RNA-seq sequence alignment, quantification, imaging, and analysis For each read, we trimmed the first 6 nucleotides and the last nucleotides at the point at which the Phred score of an examined base fell below 20 using in-house scripts.

Tissue processing for scRNA-seq, single-cell capture, library preparation, and sequencing We applied scRNA-seq using droplet microfluidics 10x Chromium on a single-cell suspension dissociated from E9. Supporting information. S1 Fig. S2 Fig. S3 Fig. S4 Fig.

Transverse histological section of E 9. S5 Fig. Uterine sections from nonpregnant or E9. S6 Fig. Single-cell RNA-seq analysis of E9. S7 Fig. Single-cell RNA-seq analysis of the proliferation status of immune and nonhematopoietic cells in the E9. S8 Fig. BMDCs do not fuse with host decidual cells. S9 Fig. Pregnancy induces mobilization of MSCs to peripheral blood. S10 Fig. Hoxa11 expression is restricted to nonhematopoietic cells. S11 Fig.

S12 Fig. S13 Fig. S14 Fig. Immune subpopulations in E9. S15 Fig. S16 Fig. Lif mRNA expression levels in the implantation site on E5. S1 Table. Primary and secondary antibodies. S2 Table. S1 Data. Underlying data. S2 Data. Four hundred ninety-eight genes commonly differentially expressed in E5. KO, knockout; WT, wild-type. Acknowledgments We would like to thank Prof. References 1.

Decidualization of the human endometrium: mechanisms, functions, and clinical perspectives. Semin Reprod Med. Cakmak H, Taylor HS. Implantation failure: molecular mechanisms and clinical treatment. Human reproduction update. Molecular cues to implantation. Endocrine reviews. A cross-talk of decidual stromal cells, trophoblast, and immune cells: a prerequisite for the success of pregnancy.

American journal of reproductive immunology. Uterine selection of human embryos at implantation. Scientific reports. Leukocyte driven-decidual angiogenesis in early pregnancy. Engraftment of bone marrow from severe combined immunodeficient SCID mice reverses the reproductive deficits in natural killer cell-deficient tg epsilon 26 mice.

J Exp Med. Developmental control point in induction of thymic cortex regulated by a subpopulation of prothymocytes. Uterine natural killer cells pace early development of mouse decidua basalis. Molecular human reproduction. Macrophages regulate corpus luteum development during embryo implantation in mice. The Journal of clinical investigation. Experimental evidence for the bone marrow origin of granulated metrial gland cells of the mouse uterus.

Cell Tissue Res. Kearns M, Lala PK. Bone marrow origin of decidual cell precursors in the pseudopregnant mouse uterus. The Journal of experimental medicine. In situ localization and characterization of bone marrow-derived cells in the decidua of normal murine pregnancy.

Biology of reproduction. Evidence that decidual cells are not derived from bone marrow. Uterine deoxyribonucleic acid synthesis during preimplantation in precursors of stromal cell differentiation during decidualization. Multi-organ, multi-lineage engraftment by a single bone marrow-derived stem cell.

Taylor HS. Endometrial cells derived from donor stem cells in bone marrow transplant recipients. Endometrial endothelial cells are derived from donor stem cells in a bone marrow transplant recipient. Human reproduction. Bone marrow-derived cells from male donors can compose endometrial glands in female transplant recipients. American journal of obstetrics and gynecology. Du H, Taylor HS. Contribution of bone marrow-derived stem cells to endometrium and endometriosis.

Stem cells. CDpositive blood cells give rise to uterine epithelial cells in mice. Stem cells and development. Experimental evidence for bone marrow as a source of nonhematopoietic endometrial stromal and epithelial compartment cells in a murine model. Dev Cell. Unique receptor repertoire in mouse uterine NK cells. J Immunol. Identification, activation, and selective in vivo ablation of mouse NK cells via NKp Expression of beta 1 integrins in human endometrial stromal and decidual cells.

The Journal of clinical endocrinology and metabolism. DBA-lectin reactivity defines natural killer cells that have homed to mouse decidua. Candeloro L, Zorn TM. Granulated and non-granulated decidual prolactin-related protein-positive decidual cells in the pregnant mouse endometrium. A map of relationships between uterine natural killer cells and progesterone receptor expressing cells during mouse pregnancy. A single-cell survey of the human first-trimester placenta and decidua.

Sci Adv. Fusion of tumour cells with bone marrow-derived cells: a unifying explanation for metastasis. Nat Rev Cancer. A Cre-Lox P recombination approach for the detection of cell fusion in vivo. J Vis Exp. Lack of a fusion requirement for development of bone marrow-derived epithelia.

Cell Stem Cell. Stem Cells. Mesenchymal stem cells can participate in ischemic neovascularization. Plast Reconstr Surg. Diabetes irreversibly depletes bone marrow-derived mesenchymal progenitor cell subpopulations. Hoxa 11 structure, extensive antisense transcription, and function in male and female fertility. Sexually dimorphic sterility phenotypes in Hoxadeficient mice.

Mechanisms of reduced fertility in Hoxa mutant mice: uterine homeosis and loss of maternal Hoxa expression. HOXA10 is expressed in response to sex steroids at the time of implantation in the human endometrium. Sex steroids mediate HOXA11 expression in the human peri-implantation endometrium.

Abnormal uterine stromal and glandular function associated with maternal reproductive defects in Hoxa null mice. Decidual cells produce a heparin-binding prolactin family cytokine with putative intrauterine regulatory actions.

J Biol Chem. Identification of target genes for a prolactin family paralog in mouse decidua. WNT4 is a key regulator of normal postnatal uterine development and progesterone signaling during embryo implantation and decidualization in the mouse. Wnt6 is essential for stromal cell proliferation during decidualization in mice. Biol Reprod. Wnt genes in the mouse uterus: potential regulation of implantation.

Uterine Msx-1 and Wnt4 signaling becomes aberrant in mice with the loss of leukemia inhibitory factor or Hoxa evidence for a novel cytokine-homeobox-Wnt signaling in implantation. Molecular endocrinology. Differential expression of microsomal prostaglandin e synthase at implantation sites and in decidual cells of mouse uterus. Alpha-2 macroglobulin controls trophoblast positioning in mouse implantation sites. Forkhead box a2 FOXA2 is essential for uterine function and fertility.

Recombineering-based dissection of flanking and paralogous Hox gene functions in mouse reproductive tracts. Adaptive changes in the transcription factor HoxA are essential for the evolution of pregnancy in mammals. A uterine decidual cell cytokine ensures pregnancy-dependent adaptations to a physiological stressor.

The endometrium as a source of mesenchymal stem cells for regenerative medicine. Msx homeobox genes critically regulate embryo implantation by controlling paracrine signaling between uterine stroma and epithelium. PLoS Genet. Blastocyst implantation depends on maternal expression of leukaemia inhibitory factor.

Foxa2 is essential for mouse endometrial gland development and fertility. Hoxa11 regulates stromal cell death and proliferation during neonatal uterine development. Angers S, Moon RT. Proximal events in Wnt signal transduction. Nature reviews Molecular cell biology. Miller C, Sassoon DA. Wnt-7a maintains appropriate uterine patterning during the development of the mouse female reproductive tract. Postnatal deletion of Wnt7a inhibits uterine gland morphogenesis and compromises adult fertility in mice.

The bone marrow-derived human mesenchymal stem cell: potential progenitor of the endometrial stromal fibroblast. Circulating endothelial progenitor cells during human pregnancy. J Clin Endocrinol Metab. Bone-marrow-derived endothelial progenitor cells contribute to vasculogenesis of pregnant mouse uterusdagger. Multipotential mesenchymal stem cells are mobilized into peripheral blood by hypoxia.

Recruitment of endogenous bone marrow mesenchymal stem cells towards injured liver. J Cell Mol Med. Estrogen-mediated endothelial progenitor cell biology and kinetics for physiological postnatal vasculogenesis. Circulation research. Cigarette smoke inhibits recruitment of bone-marrow-derived stem cells to the uterus.

Каком ищем юриста для удаленной работы на ответ, заманчиво

Инженер-технолог пищевого производства Будьте одним из первых, кто откликнулся. Будьте одним из первых, кто откликнулся. Продавец-консультант в секс-шоп 10 - 13 грн. Мы научим и поддержим вас! Отправляйте резюме или звоните телефону! Известная компания Technicolor, которая занимается производством Оценщик золота и техники , продавец 10 грн. Полный рабочий день Ломбард "Скарбниця" Проверенные контакты Активно просматривает отклики.

Водитель погрузчика 28 - 35 грн. Полный рабочий день Нерозя А. Содержание и использование автопогрузчика согласно инструкции в его паспорте. Сообщать руководству, а также делать записи в соответствующем журнале о возникновении любых неполадок, а также принимать меры к их устранению. Требования: Физическое здоровье, ответственность. Полный рабочий день Вент-Конструктив Проверенные контакты Активно просматривает отклики.

Диспетчер виробництва 8 грн. Кредитний експерт, оценщик техники и золота 8 грн. Аналитические способности. Нестандартное мышление. Анализ журналов систем безопасности, средств безопасности и данных. Опыт работы с системами и инструментами предотвращения вторжений. Кредитный эксперт, оценщик техники 12 грн. Работа Шукаю роботу журнал. In summary, our study provides evidence that BMDCs have a nonhematopoietic physiologic contribution to the decidual stroma and play an important role in implantation and pregnancy maintenance.

Importantly, nonhematopoietic BMDCs have the ability to influence the decidual molecular milieu and overcome implantation defects. Although it remains to be established to what extent these findings in the mouse translate to the situation in humans, our data raise the prospect that BMDCs dysfunction may contribute to implantation failure or pregnancy loss in women.

Hoxa11 KO mice B6. General toxicity of the treatment regimen was monitored by measuring weights of the mice and assessing their well-being daily. BMTs were performed according to the procedure described previously [ 24 ]. The morning of vaginal plug was considered E0. Successfully bred female mice were killed at various gestational time points, including E5.

Following a 3-week recovery period, different experiments were performed. For the litter size experiment, mice were harem bred with fertility-proven WT males for 1 month and monitored for weight daily. A weight increase of 4 g was considered a sign of pregnancy, at which point the pregnant dam was separated from the male and housed singly until delivery. Following delivery, the litter size and pup weights were recorded and time to delivery was calculated.

For the implantation experiment E5. Successfully bred females were killed on E5. For the resorption experiment, plug positive female mice were killed on E Mean resorption rate per pregnancy, no. Following a 3-week recovery period, transplanted mice were bred and checked for vaginal plugs daily as described above, or killed as virgins.

BM, spleen, implantation site, or nonpregnant uterus was extracted and processed as described below. Cells were then washed once in FACS buffer. Total extracted cells per mouse were counted using a hemocytometer. Appropriate unstained and antibody IgG isotype controls were used for setting compensation and determining gates. Antibodies used in flow cytometry analysis are listed in S1 Table. After 1 week, cells were passaged and maintained until the second passage P2.

P2 cells were used for subsequent flow cytometry characterization, MSC trilineage differentiation, and decidualization experiments. Fresh Chondrogenic Differentiation Medium was replaced every 2 days, with caution not to disrupt the pellet.

Another pellet was similarly processed but incubated in chondrogenic basal medium as negative control. Images were captured using a fluorescent microscope Axio Observer, Carl Zeiss. Trilineage differentiation experiments were performed in triplicate and repeated twice. Louis, MO , combination of 0. Medium was replaced every 3 days and cells were cultured for a total of 14 days. Alternatively, cells were stained for F-actin after 9 days to visualize decidual morphological changes.

All decidualization experiments were performed in triplicate and repeated three times. F-actin staining of BM MSC cells and uterine cells was performed after 9 days in culture with different decidualization treatments.

Membranes were incubated in chemiluminescence substrate and exposed to film. Five-micrometer tissue sections were mounted on slides, followed by deparaffinization and rehydration. Slides were then boiled in sodium citrate pH 6. Sections were then incubated with biotinylated secondary antibody ; Vector Laboratories, Burlingame, CA or fluorescent secondary antibody for 1 hour. Tissue sections were counterstained with hematoxylin Sigma-Aldrich, St.

Louis, MO. Images of stained sections were obtained using an Olympus BX microscope Olympus. Primary and secondary antibodies and their respective concentrations used are listed in S1 Table. For quantitation of proliferating GFP-positive and GFP-negative cells in the endometrial stromal or decidual compartments, 12 high-power confocal microscopy fields 4 high-power fields [HPFs] from each of 3 uterine sections per animal were assessed for each gestational time point.

At least 1, cells were counted per animal. For quantification of uterine stromal areas and stromal vessel luminal area, hematoxylin—eosin HE and CD31 stained photomicrographs of KO mice were analyzed using ImageJ Image analysis software Rasband, W. Total RNA was extracted from mice uterine tissues by homogenizing the tissues in 1 mL TRIzol Life Technologies, San Francisco, CA followed by the addition of uL chloroform to the lysate for phase separation by centrifugation at 13, rpm for 15 minutes.

The RNA pellet was allowed to dry and then re-dissolved in nuclease free water. Following first-strand synthesis with random primers, second-strand synthesis was performed with dUTP for generating strand-specific sequencing libraries. Libraries were sequenced on an Illumina HiSeq with parameters set for high output, single-end chemistry, and bp sequencing.

Samples were multiplexed to six samples per lane, yielding an average of 37 million reads per sample. For each read, we trimmed the first 6 nucleotides and the last nucleotides at the point at which the Phred score of an examined base fell below 20 using in-house scripts.

If, after trimming, the read was shorter than 45 bp, the whole read was discarded. Trimmed reads were mapped to the mouse reference genome mm10 with a known transcriptome index UCSC Known Gene annotation with Tophat v2. In these results, only the reads that mapped to a single genomic unique location, with a maximum of two mismatches in the anchor region of the spliced alignment, were reported.

The default settings for all other Tophat options were used. The program Cufflinks v2. Cuffdiff was used to obtain differential gene expression between the experimental groups using first-strand library type, providing gene model annotation and the genome sequence file for detection and correction of sequence-specific bias that can be caused by random hexamer during the process of library preparation.

We applied scRNA-seq using droplet microfluidics 10x Chromium on a single-cell suspension dissociated from E9. Nano-sized droplets that each contains a single cell with the bar-coded gel bead GEMs were generated using the Chromium controller 10x Genomics. Reverse transcription was performed with polyT primers containing cell-specific bar codes, unique molecular identifiers UMIs , and adaptor sequences.

All 10x libraries were sequenced in an Illumina HISeq instrument. We used the 10x Genomics Cell Ranger software v2. For graph-based clustering and differential gene expression analysis, we used Seurat 3. In Seurat, an initial filter was applied to select only the cells that had a minimum of unique transcripts; and to select only those genes that were expressed in at least 3 cells. During normalization with sctransform, we also included in the model the mitochondrial mapping percentage as an unwanted source of variation.

The expression of established lineage marker genes was used to assign cell types. Normally distributed data were analyzed using the Student unpaired two-tailed t test for the comparison of two groups, and one-way ANOVA with Tukey multiple comparison test for multiple group comparison.

If data were not normally distributed, or if distribution could not be determined due to small sample size, data were analyzed using a Mann-Whitney U test. For time to delivery Meier-Kaplan curves, differences between curves were evaluated by applying the log-rank test Kaplan-Meier log-rank value, p -value. A A schematic of the 5-FU—based submyeloablation protocol used for non-gonadotoxic BM transplantation.

B Imaging using a fluorescent camera following dissection to separate the uterus U , placenta P , and embryo E on E9. Right panel and middle panel are GFP fluorescence and X-ray images, respectively. They were analyzed by multicolor flow cytometry B. D-F Fluorescent images of trilineage differentiation of P-2 cultured BM cells grown in adipogenic media D , osteogenic media E or chondrogenic media F.

Nuclei are stained with DAPI blue. The bottom row for each panel is a higher magnification of the area in the middle row enclosed by a rectangle. Primary P-2 mouse uterine stromal cells served as positive control for decidualization. Values shown are expression levels relative to day 3. Results shown are the average of three independent experiments carried out in duplicates.

A Multicolor flow cytometry analysis of peripheral blood cells extracted from mice transplanted with BM from GFP donors following 5-FU submyeloablation. In the middle is the low-magnification image showing the mesometrial and antimesometrial sides of the implantation site. The mesometrial side is the side where the placenta, decidua basalis DB , and the major blood vessels are located.

The mesometrial lymphoid aggregate MLAp is a transient structure between the myometrial layers that surrounds the radial branches of the uterine artery. The antimesometrial side contains the rest of the maternal decidua in contact with the invading trophoblast. Red dashed line demarcates the giant cell GC layer. Maternal vascular spaces black dash have brown GFP-stained platelets black arrows and are interspersed between trophoblast cells and fetal vascular spaces green dash.

Red arrows point to nucleated red blood cells characteristic of fetal vascular spaces. Red dashed line demarcates the GC layer. Red arrows point to some decidual cells. A star demarcates the new lumen. GFP, green fluorescent protein. B Clustering of immune cells and DSCs following UMAP-based visualization of expression differences for cells using established lineage markers and C the same plot showing eGFP expression distribution within the same clusters.

Percentages shown are of cells positive for the respective marker in each group. Bottom panel shows flow cytometry of dissociated uterine implantation site cells E9. Numbers in each quadrant indicate mean percentage of cells. Multicolor flow cytometry was performed on peripheral blood and BM cells of nonpregnant and pregnant mice on E5.

Cells from BM tibia or femur , spleen, peripheral blood, or implantation site of E9. A Live single cells of nonpregnant or pregnant E9. Representative graphs are shown. Top panel shows multicolor flow cytometry gating strategy. Live leukocyte single cells were gated on CD The endometrial area in each photomicrograph is demarcated by a black dashed line.

We would like to thank Prof. Gil Mor for his helpful insights and suggestions, Kristin Milano for technical assistance with immunostaining, and Gouzel Tokmoulina for technical assistance with flow cytometry. Abstract Decidua is a transient uterine tissue shared by mammals with hemochorial placenta and is essential for pregnancy.

Eaves, B. Introduction The decidua is a transient tissue lining the uterus of mammals with hemochorial placenta mice, humans, and numerous other mammalian species and is essential for pregnancy in these species. Download: PPT. Fig 1. Spatial and temporal contribution of BMDCs to decidua throughout mouse pregnancy.

Fig 2. Characterization of uterine BMDCs throughout pregnancy. Fig 4. Hoxaexpressing BMDCs induce expression of known decidualization genes in the implantation site to rescue pregnancy The maternal component of the implantation site consists of stromal cells of the uterine endometrium that undergo a tightly controlled process of decidualization upon contact with the early embryo.

Fig 5. Fig 6. BM transplantation from WT donors leads to stromal expansion and induces glandular formation in HoxAnull mice. Fig 7. BM transplantation from WT donors leads to decidualization reaction in Hoxanull mice. Discussion It is well established that uterine implantation sites are areas of infiltration of many BM-derived immune cells, which play important roles at the maternal—fetal interface to promote successful pregnancy reviewed in [ 6 ].

Animal studies Hoxa11 KO mice B6. F-actin staining F-actin staining of BM MSC cells and uterine cells was performed after 9 days in culture with different decidualization treatments. Image quantification and analysis For quantitation of proliferating GFP-positive and GFP-negative cells in the endometrial stromal or decidual compartments, 12 high-power confocal microscopy fields 4 high-power fields [HPFs] from each of 3 uterine sections per animal were assessed for each gestational time point.

RNA-seq sequence alignment, quantification, imaging, and analysis For each read, we trimmed the first 6 nucleotides and the last nucleotides at the point at which the Phred score of an examined base fell below 20 using in-house scripts. Tissue processing for scRNA-seq, single-cell capture, library preparation, and sequencing We applied scRNA-seq using droplet microfluidics 10x Chromium on a single-cell suspension dissociated from E9.

Supporting information. S1 Fig. S2 Fig. S3 Fig. S4 Fig. Transverse histological section of E 9. S5 Fig. Uterine sections from nonpregnant or E9. S6 Fig. Single-cell RNA-seq analysis of E9. S7 Fig. Single-cell RNA-seq analysis of the proliferation status of immune and nonhematopoietic cells in the E9.

S8 Fig. BMDCs do not fuse with host decidual cells. S9 Fig. Pregnancy induces mobilization of MSCs to peripheral blood. S10 Fig. Hoxa11 expression is restricted to nonhematopoietic cells. S11 Fig. S12 Fig. S13 Fig. S14 Fig. Immune subpopulations in E9. S15 Fig. S16 Fig. Lif mRNA expression levels in the implantation site on E5.

S1 Table. Primary and secondary antibodies. S2 Table. S1 Data. Underlying data. S2 Data. Four hundred ninety-eight genes commonly differentially expressed in E5. KO, knockout; WT, wild-type. Acknowledgments We would like to thank Prof. References 1. Decidualization of the human endometrium: mechanisms, functions, and clinical perspectives. Semin Reprod Med. Cakmak H, Taylor HS. Implantation failure: molecular mechanisms and clinical treatment.

Human reproduction update. Molecular cues to implantation. Endocrine reviews. A cross-talk of decidual stromal cells, trophoblast, and immune cells: a prerequisite for the success of pregnancy. American journal of reproductive immunology. Uterine selection of human embryos at implantation. Scientific reports. Leukocyte driven-decidual angiogenesis in early pregnancy.

Engraftment of bone marrow from severe combined immunodeficient SCID mice reverses the reproductive deficits in natural killer cell-deficient tg epsilon 26 mice. J Exp Med. Developmental control point in induction of thymic cortex regulated by a subpopulation of prothymocytes. Uterine natural killer cells pace early development of mouse decidua basalis.

Molecular human reproduction. Macrophages regulate corpus luteum development during embryo implantation in mice. The Journal of clinical investigation. Experimental evidence for the bone marrow origin of granulated metrial gland cells of the mouse uterus.

Cell Tissue Res. Kearns M, Lala PK. Bone marrow origin of decidual cell precursors in the pseudopregnant mouse uterus. The Journal of experimental medicine. In situ localization and characterization of bone marrow-derived cells in the decidua of normal murine pregnancy.

Biology of reproduction. Evidence that decidual cells are not derived from bone marrow. Uterine deoxyribonucleic acid synthesis during preimplantation in precursors of stromal cell differentiation during decidualization. Multi-organ, multi-lineage engraftment by a single bone marrow-derived stem cell. Taylor HS. Endometrial cells derived from donor stem cells in bone marrow transplant recipients.

Endometrial endothelial cells are derived from donor stem cells in a bone marrow transplant recipient. Human reproduction. Bone marrow-derived cells from male donors can compose endometrial glands in female transplant recipients. American journal of obstetrics and gynecology.

Du H, Taylor HS. Contribution of bone marrow-derived stem cells to endometrium and endometriosis. Stem cells. CDpositive blood cells give rise to uterine epithelial cells in mice. Stem cells and development. Experimental evidence for bone marrow as a source of nonhematopoietic endometrial stromal and epithelial compartment cells in a murine model.

Dev Cell. Unique receptor repertoire in mouse uterine NK cells. J Immunol. Identification, activation, and selective in vivo ablation of mouse NK cells via NKp Expression of beta 1 integrins in human endometrial stromal and decidual cells. The Journal of clinical endocrinology and metabolism. DBA-lectin reactivity defines natural killer cells that have homed to mouse decidua. Candeloro L, Zorn TM. Granulated and non-granulated decidual prolactin-related protein-positive decidual cells in the pregnant mouse endometrium.

A map of relationships between uterine natural killer cells and progesterone receptor expressing cells during mouse pregnancy. A single-cell survey of the human first-trimester placenta and decidua. Sci Adv. Fusion of tumour cells with bone marrow-derived cells: a unifying explanation for metastasis. Nat Rev Cancer. A Cre-Lox P recombination approach for the detection of cell fusion in vivo. J Vis Exp. Lack of a fusion requirement for development of bone marrow-derived epithelia.

Cell Stem Cell. Stem Cells. Mesenchymal stem cells can participate in ischemic neovascularization. Plast Reconstr Surg. Diabetes irreversibly depletes bone marrow-derived mesenchymal progenitor cell subpopulations.

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